• Title/Summary/Keyword: high performance liquid chromatography-mass spectrometry (HPLC-MS)

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Separation of Tetrodotoxin, DHA and EPA from Pufferfish Liver Waste (복어간 폐기물로부터 Tetrodotoxin, DHA 및 EPA의 분리)

  • 차병윤;최진석;임정규;이동익;이원갑;이은열;김희숙;김동수
    • Journal of Life Science
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    • v.10 no.2
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    • pp.115-124
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    • 2000
  • The present study was undertaken to separate the available components effectively, such as tetrodotoxin(TTX), docosahexaenoic acid(DHA, C22:6,ω-3) and eicosapentaenoic acid (EPA, C20:5,ω -3) from pufferfish liver waste, which are known to have high values as bioactive materials. By using ultrafiltration, it was possible to separate high contents of 68mg TTX from pufferfish liver waste. In contrast, by activated charcoal column, it was to obtain about 54mg TTX. The recovering ratios were 65.3% and 45.0% in the two different methods of ultrafiltration and activated charcoal column, respectively. From the results of HPLC and gas chromatography-mass spectrometry(GC-MS), the obtained toxins were identified to be TTX and its derivatives. In addition, it was also possible to obtain 72.3g DHA and 11.4g EPA from 1kg of pufferfish liver by high performance liquid chromatography (HPLC). These amounts of DHA and EPA were also 17.70% and 1.04% in the total lipid of pufferfish liver oil from analysis of gas chromatography(GC), respectively.

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Simultaneous Determination of the Novel Neuroprotective Agent KR-31378 and its Metabolite KR-31612 Using High Performance Liquid Chromatography with Tandem Mass Spectrometry in Human Plasma

  • Kim, John;Ji, Hye-Young;Yoo, Sung-Eun;Kim, Sun-Ok;Lee, Dong-Ha;Lim, Hong;Lee, Hye-Suk
    • Archives of Pharmacal Research
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    • v.25 no.5
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    • pp.647-651
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    • 2002
  • An LC/MS/MS method for the simultaneous determination of a neuroprotective agent for ischemia-reperfusion damage, KR-31378 and its N-acetyl metabolite KR-31612 in human plasma was developed. KR-31378, KR-31612 and the internal standard. KR-31543 were extracted from human plasma by liquid-liquid extraction. A reverse-phase HPLC separation was performed on Luna phenylhexyl column with the mixture of acetonitrile-5 mM ammonium formate (55:45, v/v) as mobile phase. The detection of analytes was performed using an electrospray ionization tandem mass spectrometry in the multiple reaction monitoring mode. The lower limits of quantification for KR-31378 and KR-31612 were 2.0 ng/ml. The method showed a satisfactory sensitivity, precision, accuracy, recovery and selectivity.

Determination of mandipropamid residues in agricultural commodities using high-performance liquid chromatography with mass spectrometry (고성능액체크로마토그래피를 이용한 농산물 중 Mandipropamid의 잔류분석법 확립)

  • Kwon, Chan Hyeok;Chang, Moon Ik;Im, Moo Hyeog;Choi, Hoon;Jung, Da I;Lee, Su Chan;Yu, Jin Young;Lee, Young Deuk;Lee, Jong Ok;Hong, Moo Ki
    • Analytical Science and Technology
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    • v.21 no.6
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    • pp.518-525
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    • 2008
  • Mandipropamid is a new mandelamide-type fungicide to control foliar Oomycete pathogens in some vegetables. An analytical method was developed to determine mandipropamid residues in agricultural commodities using high-performance liquid chromatography (HPLC) and liquid chromatography/mass spectrometry (LC/MS). Mandipropamid was extracted with methanol from grape, tomato, green pepper, Chinese cabbage and potato samples. The extract was diluted with saturated sodium chloride solution and distilled water, and dichloromethane partition was followed to recover the mandipropamid from the aqueous phase. Florisil column chromatography was employed to further remove interfering co-extractives prior to HPLC analysis. Reverse-phased HPLC was successfully applied to determine mandipropamid in sample extracts with the detection at its ${\lambda}_{max}$ (223 nm). Overall recoveries of mandipropamid from fortified samples averaged $99.8{\pm}1.7$ (n=6), $89.3{\pm}5.3$ (n=6), $98.7{\pm}2.2$ (n=6), $99.7{\pm}6.8$ (n=6) and $91.1{\pm}3.1$ (n=6) for grape, tomato, green pepper, Chinese cabbage and potato, respectively. Limit of quantification of the method was 0.02~0.04 mg/kg for all samples. A LC/mass spectrometry with selected-ion monitoring was also provided to confirm the suspected residue. The proposed method was reproducible and sensitive enough to determine the terminal residue of mandipropamid in agricultural commodities.

Identification of Natural dyes used in 16th pink Dallryeong (Official's robe in Joseon Dynasty) Excavated from Cheonan, Chungnam (충남 천안시 출토 16세기 분홍 단령에 사용된 염재 동정)

  • Chae, Jeongmin;Ryu, Hyo-Seon
    • Journal of Conservation Science
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    • v.31 no.3
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    • pp.299-308
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    • 2015
  • Aim of this study is to identify dyestuff of the Dallryeong(official's robe in Joseon Dynasty, 16th century) excavated from Yuryang-dong, Cheonan, in 1996. For this purpose, extracted dyestuffs from Dallryeong fabric and from natural dyestuffs for red color(safflower, Sapanwood, Madder) which are presumed to have been used in the Dallryeong, are analyzed and compared by high performance liquid chromatography(HPLC). As a result, HPLC chromatogram of extracts of the Dallryeong's dyestuff and safflower are showed a peak at 17.5 minutes. The UV/Vis spectra of the samples are showed the maximum absorption wavelength at 519nm. This result is identical with the analysis of the previous studies on red dyestuff of safflower. In addition, the analysis of Mass Spectrometry(MS) showed the identical result of the peak with m/z 910. Following these results, excavated pink Dallryeong were considered to have been dyed with safflower.

Rapid Identification of Radical Scavenging Compounds in Blueberry Extract by HPLC Coupled to an On-line ABTS Based Assay and HPLC-ESI/MS

  • Kim, Chul-Young;Lee, Hee-Ju;Lee, Eun-Ha;Jung, Sang-Hoon;Lee, Dong-Un;Kang, Suk-Woo;Hong, Sae-Jin;Um, Byung-Hun
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.846-849
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    • 2008
  • This study employed high performance liquid chromatography (HPLC) coupled to an on-line $ABTS^+$ radical scavenging detection (RSD) system along with HPLC-electro spin impact/mass spectrometry (ESI/MS), to rapidly determine and identify antioxidant compounds occurring in blueberry extract. The extract was separated by HPLC, and then the radical scavenging activities of the separated compounds were evaluated by the on-line coupled $ABTS^+$-RSD system. The negative peaks of the $ABTS^+$-RSD system, which indicates the presence of antioxidant activity, were monitored by measuring the decrease in absorbance at 734 nm. The active components in the blueberry extract were identified by HPLC-ESI/MS using their MS spectra and retention times. According to the data acquired from the on-line HPLC-$ABTS^+$-based assay and HPLC-ESI/MS systems, the antioxidant compounds detected in the blueberry extract were identified as chlorogenic acid and 11 anthocyanins.

The separation of arsenic metabolites in urine by high performance liquid chromatography-inductively coupled plasma-mass spectrometry

  • Chung, Jin-Yong;Lim, Hyoun-Ju;Kim, Young-Jin;Song, Ki-Hoon;Kim, Byoung-Gwon;Hong, Young-Seoub
    • Environmental Analysis Health and Toxicology
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    • v.29
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    • pp.18.1-18.9
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    • 2014
  • Objectives The purpose of this study was to determine a separation method for each arsenic metabolite in urine by using a high performance liquid chromatography (HPLC)-inductively coupled plasma-mass spectrometer (ICP-MS). Methods Separation of the arsenic metabolites was conducted in urine by using a polymeric anion-exchange (Hamilton PRP X-100, $4.6mm{\times}150mm$, $5{\mu}m$) column on Agilent Technologies 1260 Infinity LC system coupled to Agilent Technologies 7700 series ICP/MS equipment using argon as the plasma gas. Results All five important arsenic metabolites in urine were separated within 16 minutes in the order of arsenobetaine, arsenite, dimethylarsinate, monomethylarsonate and arsenate with detection limits ranging from 0.15 to $0.27{\mu}g/L$ ($40{\mu}L$ injection). We used G-EQUAS No. 52, the German external quality assessment scheme and standard reference material 2669, National Institute of Standard and Technology, to validate our analyses. Conclusions The method for separation of arsenic metabolites in urine was established by using HPLC-ICP-MS. This method contributes to the evaluation of arsenic exposure, health effect assessment and other bio-monitoring studies for arsenic exposure in South Korea.

The analysis of the urushiol congeners from the extracts of lacquer trees (옻나무 추출물 중 우루시올 동종체의 함량비 분석)

  • Cho, Yumi;Jung, Yu-Kyung;Kim, Jinsan;Lee, Joonbae;Paeng, Ki-Jung
    • Analytical Science and Technology
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    • v.22 no.1
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    • pp.65-74
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    • 2009
  • Active components of lacquer tree referred to as urushiol congeners, which are catechol derivatives with various alkyl or alkenyl substituents. The olefin side chains typically have one, two or three double bonds. In this study, the each congener's ratio analysis of extracts from korean lacquer tree are compared to the one from other asian lacquer tree. Extraction was performed using liquid-liquid extraction (LLE) method with soxhlet system from tree's bark and sap. Extracts were analyzed by reverse phase liquid chromatography and on-line electro spray ionization mass spectrometry (LC-MS/MS).

Detection of Undeclared Betamethasone Derivatives in Cosmetic Products Labeled to Contain Zinc Pyrithione as the Active Ingredient (아연피리치온을 유효성분으로 표기한 화장품류에서 미표기 성분인 베타메타손 유도체의 검출)

  • Lee, Jeong-Pyo;Park, Sung-Hwan;Yang, Seong-Jun;Kim, Sun-Mi;Son, Kyung-Hun;Yun, Mi-Ok;Choi, Sang-Sook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.1
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    • pp.11-17
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    • 2009
  • Betamethasone propionate, an anti-inflammatory glucocorticosteroid, was detected in cosmetics with no indication on the label of this compound as an ingredient. The product was formulated as a topical spray or shampoo and labeled to contain zinc pyrithione as the active ingredient. A thin-layer chromatographic analysis was carried out on silica gel plates to provide a first indication about the presence of a compound with steroid structure and reactivity; then high-performance liquid chromatography (HPLC) separation allowed the identification of the corticosteroid agent and its quantification. To identify the corticosteroid agent from these commercial samples we collected the fractions suspected to have ketol steroids by prep HPLC and identified the compound as betamethasone propionate by NMR and MS spectrometry. Then we synthesized the standard for the betamethasone 17-propionate and 21-propionate and quantitate the corticosteroids from the sample by HPLC with that standards. By this method we identified the corticosteroid compounds from some commercial cosmetics such as zinc pyrithione sprays. The finding of betamethasone propionate in the products was shown by comparison to an authenticated standard of betamethasone propionate by retention time on reverse-phase HPLC. Two of the tested products contained betamethasone propionate at the levels of 0.005 ${\sim}$ 0.02% and the others were free of betamethasone propionate.

Improved HPLC-UV method for determination of five synthetic dyes in Typha orientalis

  • Ko, Kyung Yuk;Choi, Eun Young;Jeong, Se Hee;Paek, Ock jin;Lee, Chan;Heo, Huijin;Oh, She-Wook;Lee, Chulhyun;Kang, Juhye;Cho, Sooyeul
    • Analytical Science and Technology
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    • v.34 no.4
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    • pp.160-171
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    • 2021
  • Synthetic azo dyes are used extensively in herbal medicines to render the medicines more visually attractive to consumers. This study developed and validated a rapid high-performance liquid chromatography (HPLC) method to determine whether synthetic colorants such as Tartrazine, Auramine O, Metanil yellow, Sunset yellow, and Orange II are used extensively in Typha orientalis. To increase the recovery of the synthetic dyes, this method employed containing 50 mM ammonium acetate in 70 % methanol at first extraction and 100 mM HCl in 70 % methanol at second extraction. Five synthetic pigments in Typha orientalis were separated by gradient elution with a mobile phase consisting of acetonitrile and 50 mM ammonium acetate in distilled water at ultra-violet (UV) detection 428 nm or 500 nm. Additionally, this study established the liquid chromatography tandem mass spectrometry (LC-MS/MS) method to confirm positive samples suspected by HPLC results. The HPLC-UV method had good linearity, indicating r2> 0.999. The recoveries of the samples spiked with three different concentration ranged from 73.8~91.5 %, and relative standard deviation values indicated 0.2~5.2 %. The established LC-MS/MS could successfully identify the synthetic pigments in herbal medicine samples. The study demonstrates that Typha orientalis adulterated by yellowish synthetic dyes can be successfully distinguished when using the HPLC-UV method.

Anticholinesterase activity of Cinnamomum zeylanicum L. leaf extract

  • Dalai, Manoj Kumar;Bhadra, Santanu;Chaudhary, Sushil Kumar;Chanda, Joydeb;Bandyopadhyay, Arun;Mukherjee, Pulok K.
    • CELLMED
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    • v.4 no.2
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    • pp.11.1-11.6
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    • 2014
  • Cinnamomum zeylanicum (C. zeylanicum) is a tropical evergreen tree of Lauraceae family. It is one of the oldest culinary spices known and used traditionally in many cultures for centuries. In addition to its culinary uses, cinnamon also possesses as a folk remedy of many health disease condition including analgesic, antiseptic, antispasmodic, aphrodisiac, astringent, carminative, haemostatic, insecticidal, and parasiticide and memory enhancing property. This study was aimed to assess the acetylcholinesterase and butyrylcholinesterase inhibitory activity of standardized methanol extract of the C. zeylanicum. Gas chromatography - mass spectrometry (GC-MS) and high performance liquid chromatography (HPLC) analysis were done to identify the presence of eugenol as chemical component and support the neuroprotective activity in the extract. Anticholinesterase inhibitory activity of crude methanol extract of C. zeylanicum leaves and cinnamon oil were evaluated by 96-well microtiter plate assay and thin layer chromatography bioassay detection methods. This study revealed that cinnamon oil ($IC_{50}:45.88{\pm}1.94{\mu}g/ml$) has better anticholinesterase activity than methanol extract ($IC_{50}:77.78{\pm}0.03{\mu}g/ml$). In HPLC analysis, retention time of eugenol in cinnamon oil was found to be 15.81 min which was comparable with the retention time (15.99 min) of the reference standard, eugenol. Seven chemical compounds were identified by GC-MS analysis, in which eugenol as an important phytoconstituents. Thus the phytochemicals from C. zeylanicum methanol leaves extract could be developed as potential source of anticholinesterase activity, with particular benefit in the symptomatic treatment of Alzheimer's disease.